Characterization of the hatching enzyme from embryos of an anuran amphibian, Rana pirica. 1998

Y Kitamura, and C Katagiri
Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo, Japan.

The culture medium in which prehatching embryos of the frog, Rana pirica, were cultured (hatching medium) solubilized the vitelline coat (VC) of unfertilized eggs and contained molecules reactive to antibodies (anti-UVS.2) against the Xenopus hatching enzyme (HE). The hydrolyzing activity of the hatching medium against Pro-Phe-Arg-MCA was inhibited dose-dependently by the same antibodies. Using anti-UVS.2 as a probe, we purified two distinct 56 kDa molecules exhibiting Pro-Phe-Arg-MCA hydrolyzing activity. These 56 kDa molecules, which were separable on anion exchange chromatography, were the same with respect to VC solubilizing activity and a substrate specificity for various MCA-peptides, and they were regarded as charge isomers that function as the HE. The hydrolyzing activity against Pro-Phe-Arg-MCA of HE was optimal at pH of 7.6, with the apparent Km value of 250 microM at 30 degreesC. The activity was strongly inhibited by DFP and EDTA, and was accelerated by extremely low concentrations of Mg2+ and Zn2+, indicating the serine protease and metalloprotease nature of the HE. The HE was glycosylated and was present as a putative proenzyme form of 63 kDa.

UI MeSH Term Description Entries
D008666 Metalloendopeptidases ENDOPEPTIDASES which use a metal such as ZINC in the catalytic mechanism. Metallo-Endoproteinases,Metalloendopeptidase
D009842 Oligopeptides Peptides composed of between two and twelve amino acids. Oligopeptide
D010455 Peptides Members of the class of compounds composed of AMINO ACIDS joined together by peptide bonds between adjacent amino acids into linear, branched or cyclical structures. OLIGOPEPTIDES are composed of approximately 2-12 amino acids. Polypeptides are composed of approximately 13 or more amino acids. PROTEINS are considered to be larger versions of peptides that can form into complex structures such as ENZYMES and RECEPTORS. Peptide,Polypeptide,Polypeptides
D011898 Ranidae The family of true frogs of the order Anura. The family occurs worldwide except in Antarctica. Frogs, True,Rana,Frog, True,True Frog,True Frogs
D002413 Cations, Divalent Positively charged atoms, radicals or groups of atoms with a valence of plus 2, which travel to the cathode or negative pole during electrolysis. Divalent Cations
D004625 Embryo, Nonmammalian The developmental entity of a fertilized egg (ZYGOTE) in animal species other than MAMMALS. For chickens, use CHICK EMBRYO. Embryonic Structures, Nonmammalian,Embryo, Non-Mammalian,Embryonic Structures, Non-Mammalian,Nonmammalian Embryo,Nonmammalian Embryo Structures,Nonmammalian Embryonic Structures,Embryo Structure, Nonmammalian,Embryo Structures, Nonmammalian,Embryo, Non Mammalian,Embryonic Structure, Non-Mammalian,Embryonic Structure, Nonmammalian,Embryonic Structures, Non Mammalian,Embryos, Non-Mammalian,Embryos, Nonmammalian,Non-Mammalian Embryo,Non-Mammalian Embryonic Structure,Non-Mammalian Embryonic Structures,Non-Mammalian Embryos,Nonmammalian Embryo Structure,Nonmammalian Embryonic Structure,Nonmammalian Embryos,Structure, Non-Mammalian Embryonic,Structure, Nonmammalian Embryo,Structure, Nonmammalian Embryonic,Structures, Non-Mammalian Embryonic,Structures, Nonmammalian Embryo,Structures, Nonmammalian Embryonic
D004791 Enzyme Inhibitors Compounds or agents that combine with an enzyme in such a manner as to prevent the normal substrate-enzyme combination and the catalytic reaction. Enzyme Inhibitor,Inhibitor, Enzyme,Inhibitors, Enzyme
D004792 Enzyme Precursors Physiologically inactive substances that can be converted to active enzymes. Enzyme Precursor,Proenzyme,Proenzymes,Zymogen,Zymogens,Precursor, Enzyme,Precursors, Enzyme
D006031 Glycosylation The synthetic chemistry reaction or enzymatic reaction of adding carbohydrate or glycosyl groups. GLYCOSYLTRANSFERASES carry out the enzymatic glycosylation reactions. The spontaneous, non-enzymatic attachment of reducing sugars to free amino groups in proteins, lipids, or nucleic acids is called GLYCATION (see MAILLARD REACTION). Protein Glycosylation,Glycosylation, Protein
D006863 Hydrogen-Ion Concentration The normality of a solution with respect to HYDROGEN ions; H+. It is related to acidity measurements in most cases by pH pH,Concentration, Hydrogen-Ion,Concentrations, Hydrogen-Ion,Hydrogen Ion Concentration,Hydrogen-Ion Concentrations

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